Objective To investigate the feasibility of P16 immunohistochemistry combined with routine pathology in judging the residual lesion of high grade cervical intraepithelial neoplasia.Methods Patients with cervical conization for high grade cervical intrapithelial neoplasia in this hospital from January 2014 to May 2016 were chose and divided into P16 immunohistochemical detection combined with pathological diagnosis group and pathological evaluation group according to patient's motivation.Patients with residual margins were treated in accordance with the clinical guidelines and TCT was followed up for 6 months after no margin.Then sensitivity and accuracy of two group were analyzed by gold standard of follow-up results.Results 104 patients in P16 immunohistochemical detection combined with pathological diagnosis group were negative in TCT test after 6 month of following up after surgery.However,at the time of 6 months follow-up after surgery,7 patients of 112 patients have been diagnosed with positive by TCT in pathological evaluation group.The Sensitivity and accuracy in P16 immunohistochemical detection combined with pathological diagnosis group were 100％ which were higher than pathological evaluation group.Conclusion P16 immunohistochemical detection combined with conventional pathology can accurately diagnose the cervical cutting edge of conization.

To study the expression of herpes simplex virus type 2 latency-associated transcript (LAT) open reading frame 1 (ORF1) and its anti-apoptosis function induced by actinomycin D in Vero cells. The recombinant plasmid pEGFP-ORF1 was constructed and transfected into Vero cells, and the expression of ORF1 was identified by RT-PCR. The changes of Vero cells morphology induced by actinomycin D were observed by fluorescence microscopy, Hochest33258 fluorescence staining. Cells viability was evaluated by MTT assay and cells apoptosis rate was detected by flow cytometry. Double digestion and sequencing confirmed the pEGFP-ORF1 was constructed successfully, RT-PCR showed that the target gene was highly expressed in Vero cells. Hochest33258 staining reaveals that Vero cells transfected with pEGFP-ORF1 and induced apoptosis by actinomycin D had no changes in morphology. MTT assay showed that the viabilities of Vero cells transfected with recombinant plasmid pEGFP-ORF1 and induced apoptosis by actinomycin D has no statistically significant difference compared with the untreated normal control group (P > 0.05), but remarkable higher than Vero cells transfected with empty plasmid pEGFP-C2 and induced apoptosis by actinomycin D, the difference was statistically significant (P < 0.05). Flow cytometry assay shows that the cells apoptosis rate had no significant difference between pEGFP-ORF1 group and the normal group, but the cells apoptosis rate ofpEGFP-ORF1 was lower than the pEGFP-C2 group. HSV-2 LAT ORF1 gene can be expressed in Vero cells and can protect Vero cells from apoptosis induced by actinomycin D.
Animals ; Apoptosis ; physiology ; Cercopithecus aethiops ; Dactinomycin ; Herpes Simplex Virus Protein Vmw65 ; genetics ; Herpesvirus 2, Human ; genetics ; Open Reading Frames ; genetics ; Promoter Regions, Genetic ; Transcription, Genetic ; Vero Cells ; Viral Proteins ; genetics ; Virus Activation ; Virus Latency ; genetics ; physiology

Objective To study the pre-transplant serum level of anti-endothelial cell antibody(AECA)in kidney allograft recipients and its impact on the episode of acute rejection (AR) within 6 months after transplantation. Methods A total of 495 kidney allograft recipients with pre-transplant serum between December 1998 and August 2003 in our center and 40 healthy controls(negative controls)were enrolled in the study.Clinical data including AR within 6 months after transplantation were analyzed retrospectively.The serum AECA level was measured by cyto-ELISA using EA.hy926 cells as substrate,which was shown as the ratio of P (patient)/N (negative control)=(A_(petient)-A_(blank contrnal)/(A_(negative contral)-A_(blank contral).AECA was considered positive when P/N value Was greater than the average A_(negative control)value plus two times the standard deviation.Results Positive rate of AECA was 18.8%(93/495).AECA level in hemodialysis patients who had been on hemodialysis more than 12 months was 1.43±0.37,greater than those less than 12 months(1.27± 0.32,P=0.013)and those of non-dialyzed patients(1.31±0.32,P=0.029).Correlation coeffieient between AECA level and hemodialysis duration was 0.218 (P=0.018).AR incidence in AECA positive recipients was 38.7%,greater than that in AECA negative recipients (23.4%,P=0.002). Incidence of acute T cell-mediated rejection and acute antibody-mediated rejction increased significantly (P=0.035,P=0.002 respectively).Multifactor logistic regression analysis indicated that AECA positive,PRA greater than 1 0%and high CDC level were risk factors of AR with odds ratio of 2.056,1.751 and 1.764 respectively(P=0.004,0.029,0.050). Conclusions The AECA positive in pre-transplant serum indicates the elevated risk of acute allograft rejection.The AECA level increases with prolonged hemodialysis duration.

Objective To evaluate the prophylactic efficacy of compound sulfamethoxazole (SMZco)combined with ganciclovir on severe pulmonary infection in the early stage of renal transplantation. Methods Between January 2005 and January 2006,two hundred and forty renal allograft patients in our hospital were enrolled in this study.All the patients were divided into two groups.Group A(n=84)received oral SMZco combined with intravenous ganciclovir.Group B(n=156)received intravenous ganciclovir only as control.According to the time of SMZco administration,group A was divided into two subgroups:group A1(within 2weeks after transplantation,n=43)and group A2(more than 2 weeks after transplantation,n=41).All the patients were followed up for 9 months.Incidence of pulmonary infection and effects on graft function by SMZco at different time point were investigated. Results The incidence of severe pulmonary infection and mortality of infection were significantly lower in group A than those in group B (2/84 vs 16/156,P=0.027;0/2 vs 2/16,P＜0.01).There were no significant differences between two groups in terms of age,gender,warm or cold ischemia,complement dependent cytotoxieity test results,incidence of urinary infection and Scr.The incidence of elevatedScr was significantly lower in group A2 than that in group A1(15/43 vs 2/41,P＜0.01),however,all the elevated Scr returned to basal level within 1 week after SMZco was discontinued.Conclusions Oral SMZco combined with ganciclovir administration after renal transplantation is effective on preventing severe pulmonary infection and thus improves graft and recipient survival.The administration of oral SMZco initiated more than 2 weeks after transplantation is better for graft function.

BACKGROUND:There are less studies addressing whether bone marrow mesenchymal stem cells from systemic lupus erythematosus patients are different from healthy people.
OBJECTIVE:To compare the multi-differentiation capacity of bone marrow mesenchymal stem cells isolated from systemic lupus erythematosus model mice and normal control mice.
METHODS:Bone marrow mesenchymal stem cells of systemic lupus erythematosus model mice and C57BL mice were isolated and cultured fol owed by osteogenic and adipogenic differentiations, respectively. Differentiation abilities of two kinds of mouse bone marrow mesenchymal stem cells were observed.
RESULTS AND CONCLUSION:Passaged bone marrow mesenchymal stem cells from C57BL mice were long spindle-shaped and evenly distributed, while bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice showed slow growth and were relatively smal er than those from C57BL mice. After osteogenic induction, the amount of calcium salt and calcium nodules were significantly less in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice than from normal control mice. PCR detection showed that expressions of Runx2, alkaline phosphatase and osteocalcin were also significantly decreased in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice. After adipogenic induction, the number of lipid droplets in the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice was significantly less than the control group, and PCR detection also showed significantly decreased expression of adipogenic genes, including PPARγ2 and lipoprotein lipase. These findings suggest that the bone marrow mesenchymal stem cells from systemic lupus erythematosus model mice exhibit lower osteogenic and adipogenic capacities than those from normal C57BL mice, and also have the impaired multi-differentiation ability.

Objective:To explore the clinical efficacy of liver transplantation for Wilson's disease(WD).Methods:From January 1999 to November 2021, clinical data were retrospectively reviewed for 16 recipients with WD undergoing liver transplantation.There were 9 males and 7 females with an age range of 29.5(14~54)years.They were followed up by telephone, outpatient services and hospitalization.The starting point of follow-up was operation date.And recipient death was an endpoint.Postoperative survival, improvement of neuropsychiatric symptom, changes of corneal K-F ring, altered levels of liver function and serum copper-protein at Month 1 post-operation were observed.The follow-up deadline was November 24, 2021.Results:15 recipients underwent classical orthotopic liver transplantation and the other one recipient underwent living-related liver transplantation.No perioperative deaths occurred.All 16 recipients were followed up for 122(6~260)months.The 1-, 5-, and 10-year survival rates were 93.8%、85.2%and 75.8%, respectively.Among 10 recipients with corneal K-F ring positive with varying degrees after operation and was disappeared in 2 recipients at 7 and 11 months.Among 5 recipients with neuropsychiatric manifestation, 4 recipients showed ameliorative neuropsychic symptoms with varying degrees after operation and 1 recipient died.All the levels of liver function and serum copper-protien of all recipients recovered obviously in 1 month and the 1-, 5-, and 10-year post-operation.Conclusions:Classical orthotopic liver transplantation and living-related liver transplantation not only effectively improves copper metabolism of patient with WD and relieves their severe neurological manifestation, but also improves their life and prolongs survival, which is worthy of clinical promotion.

There is currently a huge worldwide demand for donor kidneys for organ transplantation. Consequently, numerous marginal donor kidneys, such as kidneys with microthrombi, are used to save patients' lives. While some studies have shown an association between the presence of microthrombi in donor kidneys and an increased risk for delayed graft function (DGF) (McCall et al., 2003; Gao et al., 2019), other studies have demonstrated that microthrombi negatively impact the rate of DGF (Batra et al., 2016; Hansen et al., 2018), but not graft survival rate (McCall et al., 2003; Batra et al., 2016; Gao et al., 2019). In contrast, Hansen et al. (2018) concluded that fibrin thrombi were not only associated with reduced graft function six months post-transplantation but also with increased graft loss within the first year of transplantation. On the other hand, Batra et al. (2016) found no significant differences in the DGF rate or one-year graft function between recipients in diffuse and focal microthrombi groups. To date, however, the overall influence of donor kidney microthrombi and the degree of influence on prognosis remain controversial, necessitating further research.
Humans ; Thrombotic Microangiopathies ; Transplantation, Homologous ; Tissue Donors ; Kidney ; Allografts